Recently, we have shown that freshly isolated cerebromicrovascular endothelial cells (EC) obtained from SJL mice with EAE in contrast to EC separated from normal brain are I-A+ and can function as antigen presenting cells. However, the expression of Ia on the surface of normal EC can be induced by gamma-interferon, (IFN gamma) and subsequently the EC are capable of presenting antigen (J Immunol 1985; 134:3100 and 1986; 137:3428). This study centered in developing and establishing EC cultures derived from microvessels of SJL mice susceptible to EAE. The dissociated endothelium was obtained from cerebral microvessels by homogenization, centrifugation, discontinous sucrose gradient (1.0-1.5 M) and exposure to dispase (0.3%). The growth and propagation of the EC depended on fibronectin presence on the surface of plastic flasks and on endothelial cell growth supplement of the medium (GIBCO 199) containing fetal calf serum (20%). Samples from each generation of cells (1-3) stained positively with antibody to factor VIII antigen and failed to stain with anti-glial fibrilary acidic protein, anti-macrophage and anti-IA. However the expression of IA+ antigen and proliferative response of these cells was induced by exposure of the cells to IFN gamma. These findings indicate that the established murine EC culture is suitable for designing a model system to investigate the interaction between the cerebral EC and immune cells in living state, which are of importance in the understanding of the pathogenesis of several neurological disorders.